Purification and characterization of latent polyphenol oxidase from truffles (Terfezia arenaria)
•Polyphenol oxidase from truffles was purified and characterized.•PPO showed best affinity for catechol as substrate and stability under wide range of pH and temperature.•Cu+2, SDS, sarkosyl and Triton X100 activate latent form of enzyme.•AL+3, Kojic acid, L-cysteine, Metabisulfite sodium and sodium...
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| Published in: | International journal of biological macromolecules Vol. 145; pp. 885 - 893 |
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| Language: | English |
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15-02-2020
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| Abstract | •Polyphenol oxidase from truffles was purified and characterized.•PPO showed best affinity for catechol as substrate and stability under wide range of pH and temperature.•Cu+2, SDS, sarkosyl and Triton X100 activate latent form of enzyme.•AL+3, Kojic acid, L-cysteine, Metabisulfite sodium and sodium fluoride were excellent PPO inhibitors.•Kojic acid amplify heat sensitivity of enzyme and reduce loss of antioxidant potential.
The polyphenol oxidase was extracted and purified from truffles (Terfezia arenaria) and it exhibited a molecular weight of 67 kDa. The truffle PPO was able to oxidize monophenolic, o-diphenolic and triphenolic substrates. Thus, the enzyme seems to be stable under wide range of pH and temperature. Best catalytic efficiency was observed for catechol as substrate (Kcat/km; 674.2 S−1 mM−1).The effect of detergents, chaotropic agents, metal ions and eleven different inhibitors on relative activity of Truffles PPO was also investigated. A latent form of enzyme was observed and its activity was stimulated using 4 mM of SDS. Likewise, the type of inhibition and the values of KI and IC50 were reported for L-cysteine, Sodium fluoride, sodium metabisulfite and kojic acid. Besides, the effect of four concentrations of kojic acid(0.05.,0.1.,0.2 and 0.3 mM) on thermal inactivation of PPO was performed in temperature range “ 60–75° C”. The use of Kojic acid increase the rate of inactivation process and disrupt enzymatic activity. Moreover, the combined effect of temperature and kojic acid prevent from enzymatic browning reaction and maintain high antioxidant activities including ABTS scavenging activity, FRAP, and total phenolic contents. |
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| AbstractList | •Polyphenol oxidase from truffles was purified and characterized.•PPO showed best affinity for catechol as substrate and stability under wide range of pH and temperature.•Cu+2, SDS, sarkosyl and Triton X100 activate latent form of enzyme.•AL+3, Kojic acid, L-cysteine, Metabisulfite sodium and sodium fluoride were excellent PPO inhibitors.•Kojic acid amplify heat sensitivity of enzyme and reduce loss of antioxidant potential.
The polyphenol oxidase was extracted and purified from truffles (Terfezia arenaria) and it exhibited a molecular weight of 67 kDa. The truffle PPO was able to oxidize monophenolic, o-diphenolic and triphenolic substrates. Thus, the enzyme seems to be stable under wide range of pH and temperature. Best catalytic efficiency was observed for catechol as substrate (Kcat/km; 674.2 S−1 mM−1).The effect of detergents, chaotropic agents, metal ions and eleven different inhibitors on relative activity of Truffles PPO was also investigated. A latent form of enzyme was observed and its activity was stimulated using 4 mM of SDS. Likewise, the type of inhibition and the values of KI and IC50 were reported for L-cysteine, Sodium fluoride, sodium metabisulfite and kojic acid. Besides, the effect of four concentrations of kojic acid(0.05.,0.1.,0.2 and 0.3 mM) on thermal inactivation of PPO was performed in temperature range “ 60–75° C”. The use of Kojic acid increase the rate of inactivation process and disrupt enzymatic activity. Moreover, the combined effect of temperature and kojic acid prevent from enzymatic browning reaction and maintain high antioxidant activities including ABTS scavenging activity, FRAP, and total phenolic contents. The polyphenol oxidase was extracted and purified from truffles (Terfezia arenaria) and it exhibited a molecular weight of 67 kDa. The truffle PPO was able to oxidize monophenolic, o-diphenolic and triphenolic substrates. Thus, the enzyme seems to be stable under wide range of pH and temperature. Best catalytic efficiency was observed for catechol as substrate (Kcat/km; 674.2 S mM ).The effect of detergents, chaotropic agents, metal ions and eleven different inhibitors on relative activity of Truffles PPO was also investigated. A latent form of enzyme was observed and its activity was stimulated using 4 mM of SDS. Likewise, the type of inhibition and the values of K and IC were reported for L-cysteine, Sodium fluoride, sodium metabisulfite and kojic acid. Besides, the effect of four concentrations of kojic acid(0.05.,0.1.,0.2 and 0.3 mM) on thermal inactivation of PPO was performed in temperature range " 60-75° C". The use of Kojic acid increase the rate of inactivation process and disrupt enzymatic activity. Moreover, the combined effect of temperature and kojic acid prevent from enzymatic browning reaction and maintain high antioxidant activities including ABTS scavenging activity, FRAP, and total phenolic contents. |
| Author | Gouzi, Hicham Benaceur, Kenza Gargouri, Ali Nouioua, Wafa Neifar, Aref Benaceur, Farouk Chaibi, Rachid Rezzoug, Asmaa Leboukh, Mourad Cabana, Hubert Berrabah, Fethi Bouazzara, Houria |
| Author_xml | – sequence: 1 givenname: Farouk surname: Benaceur fullname: Benaceur, Farouk email: f.benaceur@lagh-univ.dz organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 2 givenname: Rachid surname: Chaibi fullname: Chaibi, Rachid organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 3 givenname: Fethi surname: Berrabah fullname: Berrabah, Fethi organization: Laboratory of Exploration and Valorization of Steppic Ecosystems, Faculty of Nature and Life Sciences, University of Ziane Achour, 17000 Djelfa, Algeria – sequence: 4 givenname: Aref surname: Neifar fullname: Neifar, Aref organization: Laboratory of Molecular Biotechnology of Eukaryotes, Biotechnology Center of Sfax, University of Sfax, Tunisia – sequence: 5 givenname: Mourad surname: Leboukh fullname: Leboukh, Mourad organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 6 givenname: Kenza surname: Benaceur fullname: Benaceur, Kenza organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 7 givenname: Wafa surname: Nouioua fullname: Nouioua, Wafa organization: Department of English, Faculty of Letters and Languages, University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 8 givenname: Asmaa surname: Rezzoug fullname: Rezzoug, Asmaa organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 9 givenname: Houria surname: Bouazzara fullname: Bouazzara, Houria organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 10 givenname: Hicham surname: Gouzi fullname: Gouzi, Hicham email: h.gouzi@lagh-univ.dz organization: Laboratory of Biological and Agricultural Sciences (LSBA), University of Amar Telidji (UATL), 03000 Laghouat, Algeria – sequence: 11 givenname: Hubert surname: Cabana fullname: Cabana, Hubert organization: Department of Civil Engineering, Université de Sherbrooke, 2500 Boulevard de l’Université, Sherbrooke, Québec J1K 2R1, Canada – sequence: 12 givenname: Ali surname: Gargouri fullname: Gargouri, Ali organization: Laboratory of Molecular Biotechnology of Eukaryotes, Biotechnology Center of Sfax, University of Sfax, Tunisia |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31778703$$D View this record in MEDLINE/PubMed |
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| Keywords | Purification, characterization Truffles kojic acid.enzymatic browning Polyphenol oxidase |
| Language | English |
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| Snippet | •Polyphenol oxidase from truffles was purified and characterized.•PPO showed best affinity for catechol as substrate and stability under wide range of pH and... The polyphenol oxidase was extracted and purified from truffles (Terfezia arenaria) and it exhibited a molecular weight of 67 kDa. The truffle PPO was able to... |
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| SubjectTerms | kojic acid.enzymatic browning Polyphenol oxidase Purification, characterization Truffles |
| Title | Purification and characterization of latent polyphenol oxidase from truffles (Terfezia arenaria) |
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